Objective To investigate the effect of different explants on emodin contents in the callus of Polygouum cuspidatum by reverse-phase HPLC. Methods The leafs,stems and leaf stalks of Polygouum cuspidatum were used as explants and induce the callus by MS medium containing 6-BA 0.2 mg/L,KT 0.2 mg/Land 2,4-D 0.5 mg/L. The contents of emodin were analyzed by HPLC. The separation was performed on Diamonsil-C18 (250 mm?4.6 mm,5 ?m) column,using methanol-0.1% H3PO4 (80∶20) as mobile phase,detected at 254 nm. The flow rate was at 1.0 mL/min,the column temperature was 27 ℃. Results The contents of emodin were 0.709 6,1.680 0 and 0.166 8 mg/g in the callus induced by leafs,stems and leaf stalks. The linear range of emodin was 0.25～5 ?g/mL,r=0.999 85,the average recovery was 101.19% and RSD was 1.20%. Conclusion HPLC method could be used for the contents determination of emodin in Polygouum cuspidatum callus and the contents of emodin was high in the callus induced by stems.

Objective To explore the role of receptor‐interacting protein kinase 3 (RIP3)‐mediated necroptosis in ulcerative colitis (UC ) . Methods The colonic mucosa tissues of eighty‐five patients diagnosed with UC were collected .Disease staging and pathological classification of UC patients were evaluated according to Mayo standard and Truelove standard ,respectively .The expressions of RIP3 and tumor necrosis factor (TNF)‐α were detected by immunohistochemistry .The cell inflammation model in HT‐29 cells was induced by interferon (IFN)‐γ.The expression of RIP3 mRNA and interleukin (IL)‐1βmRNA was determined by real time polymerase chain reaction (RT‐PCR) .Twenty healthy individuals were studied as controls .A single factor analysis of variance was performed in the comparison among groups ,and Pearson correlation analysis was used for correlation analysis .Results Among 85 patients with UC , 27 cases were mild ,30 cases were moderate and 28 cases were severe;for pathological classification ,26 were gradeⅠ ,31 were gradeⅡ and 28 were grade Ⅲ .The Mayo score was positively correlated with pathological grade (r=0 .997 , P<0 .05) .The expression of RIP3 and TNF‐α in colonic mucosa tissues of UC group increased along with the pathological classification grade .The expression of RIP3 of gradeⅠto Ⅲ was 1 .81 ± 0 .98 ,2 .77 ± 1 .26 and 4 .86 ± 2 .77 ,respectively ,and the differences were statistically significant (F=26 .10 ,P<0 .05) .The expression of TNF‐αof gradeⅠto Ⅲ was 1 .35 ± 0 .69 ,2 .61 ± 1 .41 and 4 .43 ± 2 .17 ,respectively ,and the differences were statistically significant (F=31 .80 ,P<0 .05) .The expression of IL‐1β mRNA of IFN‐γ group ,IFN‐γ+ z‐VAD group and IFN‐γ+z‐VAD+ TNF‐αgroup was 0 .68 ± 0 .27 ,1 .47 ± 0 .12 and 1 .86 ± 0 .16 ,respectively ,and the differences were statistically significant (F=38 .45 , P<0 .05) .The expression of RIP3 mRNA of IFN‐γgroup ,IFN‐γ+z‐VAD group and IFN‐γ+z‐VAD+TNF‐αgroup was 0 .46 ± 0 .13 ,1 .21 ± 0 .29 and 2 .06 ± 0 .20 ,respectively , and the differences were statistically significant (F= 55 .15 , P< 0 .05) .Conclusions TNF‐α/RIP3 signal pathway is up‐regulated in UC .RIP3‐mediated necroptosis promotes the death of colonic epithelial cells and exacerbates inflammatory response .RIP3‐mediated necroptosis may be an important mechanism in UC .

[Objective]To observe the effect of sciatic nerve and femoral nerve resection on heterotopic osteogenesis induced by recombined human bone morphogenetic protein-2(rhBMP-2) and discuss role of never innervation on bone regeneration and partial reasons for larger callus after bone fracture accompanying denervation.[Method]A total of 36 male ICR mice were divided into experimental group and control group at random.0.125mg rhBMP-2 /collagen composites were implanted into the right thigh muscle pouches after sciatic nerve and femoral nerve section in the experimental group and after sham operation in the control group.On the 7~(th),14~(th) and 21~(st) day after implantation,roentgenographic,biochemical,histological analyses and osteoclasts TRAP staining were performed to detect the effects of sciatic nerve and femoral nerve resection on bone growth initiated by rhBMP-2.[Result]On the 7~(th),14~(th) and 21~(st) day,wet weight of new bony tissues of the experimental group was obviously greater than that of the control group.Radiography showed range of bone formation in the experimental group was larger than that in the control,but density of new bone was lower than that of the latter.Biochemical detection showed value of AKP of the experimental group was significantly higher than that of the control on the 7~(th) day,while the content of Ca of the experimental group was higher than that of the control on the 14~(th) day and the content of Ca and P of the experimental group was lower than that of the control on the 21~(st) day.Histological observation showed area of new bone of the experimental group was greater than that of the control,but trabecular bone of the former was sparser than that of the latter.Image analysis of bony tissue showed the relative number of osteoclasts in the experimental group became higher on the 21~(st) day,while volume density and width of bone trabecula in the experimental group were lower than that of the control group.TRAP staining showed osteoclasts in the new bone of the experimental group were more activated than that of the control.[Conclusion]Nerve section results in enhancement of ability of bone regeneration by BMP in the early period,but trabecular bone becomes sparse in the middle and later period because of bone resorption by osteoclasts,which indicates nerve in nervation influences bone regemeration though direct and/or indirect way.

[Objective]To identify the expression of neurotrophic factors(NTFs)and their high affinity receptors(Trk)during the bone induction by recombined human bone morphogenetic protein-2(rhBMP-2)and discuss possible roles of NTFs on the osteoinductive activity of BMP.[Method]After a total of 36 ICR mice were divided into experimental group and control group at random,rhBMP-2/collagen sponge and collagen sponge were implanted into the right thigh muscle pouches of two groups respectively.Tissues in the implanted sites of two groups were removed on the 7~(th),14~(th) and 21~(st) day after implantation.Histological,immunohistochemical and RT-PCR analyses were performed to detect osteoinductive effect of rhBMP-2 and the expression of NTFs and Trk.[Result]RhBMP-2/collagen sponge displayed a potent ability inducing bone formation,while the expression of NTFs and Trk was observed in the course of osteoinduction by rhBMP-2.Especially on the 7~(th) day in the experimental group,NGF and TrkA positive immunostaining reached the peak in the stage of chondrogenesis,including chondroblasts,chondrocytes,hypertrophic chondrocytes and osteoblasts,then decreased in the number of positive cells and the intensity of immunostaining on the 14~(th) and 21~(st) day.BDNF was only observed in chondrocytes and cartilage matrix,and TrkB was expressed in chondroblasts and chondrocytes at the 7~(th) day after implantation.Expression of NT-3 was similar to that of NGF,while TrkC was expressed in chondroblasts and chondrocytes on day 7.Expression level of the mRNA of NTFs peaked at 7 days after the implantation,then decreased,which was nearly coincident with immunohistochemical results.[Conclusion]Many different kinds of cells are induced by rhBMP-2 express NTFs and Trk,which suggests that NTFs may play an important role in the osteogenesis initiated by BMP through direct and indirect pathways.

BACKGROUND:Carboxymethyl chitosan (CMCS),a chitin derivative,has a good application performance that makes it become a safe and effective hemostatic material.OBJECTIVE:To determine the hemostatic effect of CMCS and its biosecurity properties.METHODS:(1) CMCS powder was scattered on the caudal vein and liver wounds of Sprague-Dawley rats,and the hemostatic time was recorded as experimental group,while the time for natural haemostasis of the wound was recorded as control group.(2) CMCS powder was scattered on the tail,femoral artery and liver wounds of ICR mice,and the hemostatic time was recorded as experimental group,while the time for natural haemostasis of the wound was recorded as control group.(3) CMCS,Sodium dodecyl sulfate solution and distilled water were respectively applied on the skin of albino rabbits in a skin irritation test.(4) A delayed-type hypersensitivity test of CMCS was carried out by intradermal injection of CMCS in guinea pigs.(5) An intradermal irritation test was carried out by subcutaneous injection of normal saline containing CMCS and normal saline,respectively.Another intradermal irritation test was carried out by subcutaneous injection of the supernatant of CMCS olive oil extract and olive oil,respectively.RESULTS AND CONCLUSION:(1) Compared with the control group,the hemostatic time for caudal vein and liver wounds were significantly shortened in the Sprague-Dawley rats in the experimental group (P ＜ 0.01).(2) Compared with the control group,the time of hemostasis on the tail,femoral artery and liver wounds was significantly shortened in the ICR mice in the experimental group (P ＜ 0.05 or P ＜ 0.01).(3) The CMCS had no irritation to the skin of albino rabbits and no allergic reaction to the skin of guinea pigs.To conclude,the CMCS has good hemostatic effect on the wound in Sprague-Dawley rats and ICR mice,and has no skin irritation,allergic reactions and intradermal irritation reactions in albino rabbits and guinea pigs,which is a relatively safe hemostatic material.

Objective To investigate the application value of serum tumor necrosis factor alpha (TNF-α) expression in diagnosing colorectal cancer (CRC).Methods One hundred and thirty inpatients with CRC in our hospital from February 2015 to November 2016 were selected and divided into 90 cases of CRC confirmed by operation and histopathology(CRC group),including intestinal cancer with and without metastasis,and 40 cases of benign colorectal diseases(colonrectal benign disease group),at the same time 40 persons undergoing physical examination were selected as the healthy control group.The levels of TNF-α,carcinoembryonic antigen (CEA) and carbohydrate antigen (CA) 724 were collected in all subjects.The data of various groups were not normally distributed,and the nonparametric statistical method was used to analyze and compare.Results Serum TNF-α level in clinical TNM stage Ⅱ(median 9.47 pg/mL),Ⅲ(9.95 pg/mL) and Ⅳ(9.67 pg/mL) of the CRC group was significantly higher than that in TNM stage Ⅰ(6.59 pg/mL);which in the CRC with metastasis group (9.95 pg/mL) was higher than 8.03 pg/mL in CRC without metastasis group,the difference was statistically significant (P<0.05);the TNF-α expression had no obvious difference among different ages and between different genders(P>0.05).The area under the ROC curve in each test index:TNF-α was 0.868,CEA was 0.912,CA724 was 0.523,and which of their joint detection was 0.940.Conclusion The expression of TNF-α is increased in the occurrence and development process of CRC,its serum level detection has certain clinical application value,the TNF-α expression has no obvious difference in the subjects with different ages and different sexes and has wide suitability.The combine detection of serum TNF-α,CEA and CA724 in the patients with CRC significantly increases the level,the combined detection of 3-index can have complementary advantages to increase the sensitivity of CRC diagnosis,which serve as the significant indexes in CRC early diagnosis,therapeutic effect evaluation and prognosis monitoring.

Objective To study the influence of different positions in the isokinetic muscle test of knees by CON-TREX Biomechanical Test and Training System, so as to select the suitable conditions for forensic identification of muscle strength test. Methods Fifty-two healthy volunteers joined the isokinetic muscle strength test in unfixed and fixed position, respectively and in two kinds of angular speed (60°/s and 30°/s). The differences of peak torque (PT) and peak torque angle (PTA) between bilateral knee flexor and extensor were statistically analyzed. Results In the unfixed position, under the two speed, there was statistically significant difference in PT between bilateral knee flexor and extensor (P<0.05);while in the fixed position, under the two speed, there was no statistically significant difference (P>0.05). In any kind of conditions, the PTA of bilateral knee flexor and extensor did not have statistically signifi-cant difference(P>0.05). Conclusion The position of the subject influences the results of PT. So the po-sition of subject in knees isokinetic muscle test should be regulated.

Objective To study the application of isokinetic muscle testing in identification of the faked paralysis to provide scientific data for establishing a standard systemof muscle strength in forensic medicine identification. Methods Fifty-seven patients with bone fracture or nerve damage as damaged group and 128 normal subjects pretended paralysis as faked paralyzed group were included in this study. Isokinetic muscle testing was performed on bilateral knees of all subjects in the two groups. The peak torque (PT) and peak torque angle (PTA ) were compared between both sides in each group. The fea-tures of torque-time graph of two groups were classified. Results In the damaged group, the differences of PTbetween two sides of flexors and extensors were statistically significant (P<0.05), while the dif-ferences of PTA were not statistically significant (P>0.05). In faked paralyzed group, the differences of PTand PTA between two sides of flexors and extensors were both statistically significant (P<0.05). The torque-time graph of damaged knee presented mostly as single lead peak, while torque-time graph of the faked paralyzed knee presented mostly as multiple peaks. Conclusion The feature of torque-time graph could be useful to identify the faked paralyzed extremities in forensic authentication.

Objective To evaluate the application value with low-dose CT scan and quantitative analysis in airway disease of chronic obstructive pulmonary disease(COPD).Methods All subjects(34 control cases,88 cases of stable COPD patients)underwent low-dose CT chest scans,and 88 COPD patients underwent pulmonary function tests.Measuring the airway dimensions of 3th generation in apical bronchus of the right upper lobe,the posterior basal bronchus of the right lower lobe and left lower lobe.Results COPD patients with moderate to extremely severe airflow limitation demonstrated higher WA% and Pi10 in comparison with normal volunteers(all P<0.05),meanwhile patients with extremely severe airflow limitation demonstrated higher WA % and Pi10 in comparison with patients with mild airflow limitation(both P <0.05).COPD patients with severe airflow limitation demonstrated higher T/BSA and WA/BSA in comparison with normal volunteers and patients with moderate airflow limitation (all P<0.05).The WA % and Pi10 of COPD patients group were negatively correlated with ventilation indices and small airway indices(all P <0.01).Pi10 were positively correlated with lung volume indices (all P < 0.05 ).Conclusion Low-dose CT scan and quantitative analysis can provide an objective and quantitative method to evaluate the information about airway disease of COPD,inwich both WA % and Pi10 are objective indices.With the increase of air flow limitation,the bronchial wall thickness gradually increased.